This K22 Award project is designed to support the PI's transition to becoming an independent researcher at an academic institution. The PI has prepared for this research through studies in molecular biology biochemistry, microbiology, and immunology. His most recent work has shown the strong effects of soluble, multimeric CD40L- and GITRL-based molecular adjuvants on HIV DNA vaccines in mice. Using support from this K22 award, the PI would move these molecular adjuvants into adenoviral vaccines against HIV, in order to make these promising vaccines even stronger. To this end, four Specific Aims are proposed: (1) To prepare adenoviral vectors containing HIV Gag along with novel soluble multimeric CD40L or GITRL. (2) To determine if these molecularly adjuvanted adenoviruses are effective activators of dendritic cells in vitro. (3) To determine if these TNFSF-adjuvanted adenoviral vaccines induce stronger, broader, and longer-lasting responses than do their unadjuvanted counterparts. (4) To determine if these TNFSF-adjuvanted adenoviral vaccines are capable of inducing immune responses in mice that have high levels of neutralizing anti-adenoviral antibodies. These studies, when complete, promise to improve adenoviral vaccines that are already being tested in human trials against HIV. In addition, adenoviral vaccines may protect against many other infections, including influenza, SARS, Ebola, and agents of biodefense interest. Thus, this approach toward making stronger vaccines, which is the PI's scientific focus, could lead to better control of infectious diseases and has broad public health significance.